The long-term goal of our studies is to understand the role of biliary epithelium in the repair and regeneration of the liver after damage. Inherited cholangiopathies with identified genetic defects serve as model diseases to elucidate the fundamental pathophysiological mechanisms. The previously funded proposal focused on polycystic liver disease associated to Adult Dominant Polycystic Kidney Disease (PLD-ADPKD) as a paradigm for the role of angiogenic signaling in biliary diseases. We uncovered mechanisms that may be relevant for the pathogenesis of other congenital and acquired liver diseases. In fact, using mice models with inducible defects of polycystins, we found that: 1) the cystic epithelium produces VEGF and expresses its cognate receptor VEGFR2; 2) VEGF-mediated stimulation of VEGFR2 results in increased ERK1/2-dependent proliferation of the cystic epithelium, 3) PC2-defective cystic cholangiocytes, altered cellular Ca2+ homeostasis and a cAMP- dependent increase in PKA/Ras/Raf/ERK signaling results in mTOR/HIF-1-mediated stimulation of VEGF production, 4) in response to stimuli able to deplete ER Ca2+ stores, PC2 participates in store-operated Ca2+ entry (SOCE); 5) if PC2 is defective, an alternative pathway is activated (store-operated cAMP production - SOcAMP), leading to an inappropriate overproduction of cAMP; 6) VEGF/VEGFR2 play a key role on cyst growth and expansion through paracrine effects on pericystic vascular cells, and autocrine stimulation of the cystic epithelium proliferation. These findings are the basis of this new proposal which main hypothesis is that the mechanism linking PC2 to VEGF secretion and VEGFR2 expression identified in PLD-ADPKD is of general relevance in biliary pathophysiology. We will address this hypothesis through three specific aims: 1) to better understand the interactions between PC2 function store-operated Ca2+ entry and inappropriate production of cAMP, 2) to study if PC2 expression in WT cholangiocytes can be modulated by cell stressors, thereby reproducing the changes seen in PC2-defective cells; 3) to study the mechanisms leading to VEGFR2 expression in cystic and reactive cholangiocytes, and to elucidate the role of VEGF in the branching morphogenesis of the biliary epithelium during liver repair. These studies will address the novel idea that PC2 play a pivotal role in the regulation of cholangiocyte response to biliary damage acquired cholangiopathies, and that VEGF secreted by reactive cholangiocytes is a major factor in liver repair. Furthermore, our studies will increase understanding of VEGF/VEGFR2 signaling in epithelia and will address a fundamental mechanism in congenital and acquired cholangiopathies. Understanding the pathophysiology of cholangiopathies is a fundamental step for preserving liver function and prolonging the survival of patients